Rhinacanthin-C Extracted from Rhinacanthus nasutus (L.) Inhibits Cholangiocarcinoma Cell Migration and Invasion by Decreasing MMP-2, uPA, FAK and MAPK Pathways

Document Type: Research Articles

Authors

1 Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.

2 Center of Excellence for Innovation in Chemistry, Cholangiocarcinoma Research Institute, Cholangiocarcinoma Screening and Care Program (CASCAP), Faculty of Medicine, Khon Kaen University, Khon Kaen,Thailand.

3 Center of Excellence for Innovation in Chemistry, Liver Fluke and Cholangiocarcinoma Research Institute, Cholangiocarcinoma Screening and Care Program (CASCAP), Faculty of Medicine, Khon Kaen University, Khon Kaen,

4 Natural Products Research Section, Research Division, National Cancer Institute; Bangkok, Thailand.

Abstract

Cholangiocarcinoma is a malignant tumor with high metastatic and mortality rates. We investigated the effects
of rhinacanthin-C on cell proliferation, migration, invasion and the expression of proteins regulating cancer cell
invasion-regulated proteins in a cholangiocarcinoma (KKU-M156) cell line. Cytotoxicity of rhinacanthin-C was
determined by the SRB assay. Using wound-migration, chamber-migration and chamber-invasion assays, we assessed
the effects of rhinacanthin-C against KKU-M156 cells. The activities of matrix metalloproteinases 2 and 9 (MMP-2,
MMP-9) and urokinase-type plasminogen activator (uPA) were determined using gelatinase and uPA zymography
assays. The expression of invasion-regulated proteins was investigated using western-blot analysis. After treatment
with rhinacanthin-C, KKU-M156 cells exhibited antiproliferative effects in a dose-dependent manner with greater
efficacy than in Vero cells: IC50 values were 1.50 and 2.37 μM, respectively. Rhinacanthin-C significantly inhibited cell
migration and invasion of KKU-M156 cells in a dose-dependent manner. Consistent with this observation, treatment
with rhinacanthin-C was associated with a decrease in the expression levels of FAK, p-FAK, MMP-2, and a decrease in
the levels of p38-, JNK1/2- and ERK1/2-MAPK pathways as well as inhibiting NF-κB/p65 expression and translocation
of NF-κB/p65 to the nucleus. We have shown for the first time that the anti-metastatic effects of rhinacanthin-C on
KKU-M156 cells are mediated via inhibition of the expression of invasion-regulated proteins. Rhinacanthin-C may
deserve consideration as a potential agent for the treatment of cholangiocarcinoma.

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